Ciro Isidoro received his doctoral degree (Doctor of Science Phone: 0321660507 Address: Università del Piemonte Orientale, Department of Health Sciences, Via P. Solaroli 17, 28100 Novara
Keloids are characterized by increased deposition of fibrous tissue in the skin and subcutaneous ... more Keloids are characterized by increased deposition of fibrous tissue in the skin and subcutaneous tissue following an abnormal wound healing process. Although keloid etiology is yet to be fully understood, fibroblasts are known to be key players in its development. Here we analyze the antifibrotic mechanisms of Halofuginone (HF), a drug reportedly able to inhibit the TGF-β1-Smad3 pathway and to attenuate collagen synthesis, in an in-vitro keloid model using patient-derived Keloid Fibroblasts (KFs) isolated from fibrotic tissue collected during the "Scar Wars" clinical study (NCT NCT03312166). TGF-β1 was used as a pro-fibrotic agent to stimulate fibroblasts response under HF treatment. The fibrotic related properties of KFs, including survival, migration, proliferation, myofibroblasts conversion, ECM synthesis and remodeling, were investigated in 2D and 3D cultures. HF at 50 nM concentration impaired KFs proliferation, and decreased TGF-β1-induced expression of α-SMA and type I procollagen production. HF treatment also reduced KFs migration, prevented matrix contraction and increased the metallo-proteases/inhibitors (MMP/TIMP) ratio. Overall, HF elicits an anti-fibrotic contrasting the TGF-β1 stimulation of KFs, thus supporting its therapeutic use for keloid prevention and management.
Gli Autori presentano i risultati di un'attivita di ricerca e sviluppo finalizzata alla defin... more Gli Autori presentano i risultati di un'attivita di ricerca e sviluppo finalizzata alla definizione di un approccio innovativo alla progettazione ergonomica dei sistemi produttivi di beni e servizi (industria e terziario, settore pubblico e privato). L'attivita si e svolta nell'ambito del progetto "Modelli di gestione dell'ergonomia per la sicurezza sui luoghi di lavoro" finanziato dalla Regione Piemonte ed e stata orientata all'impostazione di un sistema informativo e formativo di riferimento in ambito territoriale. Nucleo fondante del progetto e stato l'assunto, fatto proprio dai decisori regionali, che l'Ergonomia nei luoghi di lavoro non rappresenta unicamente uno strumento per tutelare la sicurezza ed il benessere dei lavoratori, ma anche un valido strumento di innovazione e progresso dei sistemi produttivi. I principali risultati del lavoro svolto nella prima fase del progetto (conclusasi nel febbraio 2010) sono stati resi disponibili in un sito internet pubblico (www.ergonomia.corep.it). Il lavoro di gruppo ha portato, tra l'altro, alla definizione di una check-list di controllo in fase progettuale e alla realizzazione di un prototipo di strumento per l'analisi comparata delle piu diffuse metodologie di valutazione ergonomica (Toolkit)
Collection of Czechoslovak Chemical Communications, Oct 16, 2014
ABSTRACT NaGdF4 nanoparticles (NPs) coated with organic ligands are known to provide efficient “p... more ABSTRACT NaGdF4 nanoparticles (NPs) coated with organic ligands are known to provide efficient “positive” contrast in magnetic resonance imaging (MRI). Strongly chelating ligands, such as ethylenediaminetetraacetic acid (EDTA) and d,l-1,2-diaminopropionic-N,N,N′,N′-tetraacetic acid (EDTACOOH), which is an EDTA derivative with a functionalisable carboxylic group, were used to coat NaGdF4. The carboxylic group was used to insert a polyethylene glycol (PEG) functionality (to give EDTAPEG) to favour better suspension and stealth and/or a fluorescent dye to obtain dual optical/MRI probes. The relaxometric behaviour of these NPs was investigated as a function of the magnetic field strength, and a significant contribution from water molecules hydrogen bonded to the organic coating was evidenced. The chemical stability of the NPs was evaluated both in a physiological medium and in the presence of a strong chelating agent. Finally, the dye-functionalised NPs were tested in ovarian carcinoma cells as dual optical and MRI probes.
Neuroblastoma is the most common type of cancer in infants. In children this tumor is particularl... more Neuroblastoma is the most common type of cancer in infants. In children this tumor is particularly aggressive; despite various new therapeutic approaches, it is associated with poor prognosis. Given the importance of endosomal-lysosomal proteolysis in cellular metabolism, we hypothesized that inhibition of lysosomal protease would impact negatively on neuroblastoma cell survival. Treatment with E-64 or CA074Me (2 specific inhibitors of cathepsin B) or with pepstatin A (a specific inhibitor of cathepsin D) was cytotoxic for 2 neuroblastoma cell lines having different degrees of malignancy. Cell death was associated with condensation and fragmentation of chromatin and externalization of plasma membrane phosphatidylserine, 2 hallmarks of apoptosis. Concomitant inhibition of the caspase cascade protected neuroblastoma cells from cathepsin inhibitor-induced cytotoxicity. These data indicate that prolonged inhibition of the lysosomal proteolytic pathway is incompatible with cell survival, leading to apoptosis of neuroblastoma cells, and that the cathepsin-mediated and caspase-mediated proteolytic systems are connected and cooperate in the regulation of such an event. Since modern antitumor chemotherapy is aimed at restoring the normal rate of apoptosis in neoplastic tissues, the demonstration that endosomal-lysosomal cathepsins are involved in this process may constitute a basis for novel strategies that include cathepsin inhibitors in the therapeutic regimen.
Hydrogen peroxide, the major oxidoradical species in the central nervous system, has been involve... more Hydrogen peroxide, the major oxidoradical species in the central nervous system, has been involved in neuronal cell death and associated neurodegenerative diseases. In this study, we have investigated the involvement of the lysosomal pathway in the cytotoxic mechanism of hydrogen peroxide in human neuroblastoma cells. Alteration of lysosomal and mitochondrial membrane integrity was shown to be an early event in the lethal cascade triggered by oxidative stress. Desferrioxamine (DFO), an iron chelator that abolishes the formation of reactive oxygen species within lysosomes, prevented lysosome leakage, mitochondrial permeabilization and caspase-dependent apoptosis in hydrogen peroxide-treated cells. Inhibition of cathepsin D, not of cathepsin B, as well as small-interference RNA-mediated silencing of the cathepsin D gene prevented hydrogen peroxide-induced injury of mitochondria, caspase activation, and TUNEL-positive cell death. Cathepsin D activity was shown indispensable for translocation of Bax onto mitochondrial membrane associated with oxidative stress. DFO abolished both the cytosolic relocation of Cathepsin D and the mitochondrial relocation of Bax in hydrogen peroxide-treated cells. siRNA-mediated down-regulation of Bax expression protected the cells from oxidoradical injury. The present study identifies the lysosome as the primary target and the axis cathepsin D-Bax as the effective pathway of hydrogen peroxide lethal activity in neuroblastoma cells.
Keloids are characterized by increased deposition of fibrous tissue in the skin and subcutaneous ... more Keloids are characterized by increased deposition of fibrous tissue in the skin and subcutaneous tissue following an abnormal wound healing process. Although keloid etiology is yet to be fully understood, fibroblasts are known to be key players in its development. Here we analyze the antifibrotic mechanisms of Halofuginone (HF), a drug reportedly able to inhibit the TGF-β1-Smad3 pathway and to attenuate collagen synthesis, in an in-vitro keloid model using patient-derived Keloid Fibroblasts (KFs) isolated from fibrotic tissue collected during the "Scar Wars" clinical study (NCT NCT03312166). TGF-β1 was used as a pro-fibrotic agent to stimulate fibroblasts response under HF treatment. The fibrotic related properties of KFs, including survival, migration, proliferation, myofibroblasts conversion, ECM synthesis and remodeling, were investigated in 2D and 3D cultures. HF at 50 nM concentration impaired KFs proliferation, and decreased TGF-β1-induced expression of α-SMA and type I procollagen production. HF treatment also reduced KFs migration, prevented matrix contraction and increased the metallo-proteases/inhibitors (MMP/TIMP) ratio. Overall, HF elicits an anti-fibrotic contrasting the TGF-β1 stimulation of KFs, thus supporting its therapeutic use for keloid prevention and management.
Gli Autori presentano i risultati di un'attivita di ricerca e sviluppo finalizzata alla defin... more Gli Autori presentano i risultati di un'attivita di ricerca e sviluppo finalizzata alla definizione di un approccio innovativo alla progettazione ergonomica dei sistemi produttivi di beni e servizi (industria e terziario, settore pubblico e privato). L'attivita si e svolta nell'ambito del progetto "Modelli di gestione dell'ergonomia per la sicurezza sui luoghi di lavoro" finanziato dalla Regione Piemonte ed e stata orientata all'impostazione di un sistema informativo e formativo di riferimento in ambito territoriale. Nucleo fondante del progetto e stato l'assunto, fatto proprio dai decisori regionali, che l'Ergonomia nei luoghi di lavoro non rappresenta unicamente uno strumento per tutelare la sicurezza ed il benessere dei lavoratori, ma anche un valido strumento di innovazione e progresso dei sistemi produttivi. I principali risultati del lavoro svolto nella prima fase del progetto (conclusasi nel febbraio 2010) sono stati resi disponibili in un sito internet pubblico (www.ergonomia.corep.it). Il lavoro di gruppo ha portato, tra l'altro, alla definizione di una check-list di controllo in fase progettuale e alla realizzazione di un prototipo di strumento per l'analisi comparata delle piu diffuse metodologie di valutazione ergonomica (Toolkit)
Collection of Czechoslovak Chemical Communications, Oct 16, 2014
ABSTRACT NaGdF4 nanoparticles (NPs) coated with organic ligands are known to provide efficient “p... more ABSTRACT NaGdF4 nanoparticles (NPs) coated with organic ligands are known to provide efficient “positive” contrast in magnetic resonance imaging (MRI). Strongly chelating ligands, such as ethylenediaminetetraacetic acid (EDTA) and d,l-1,2-diaminopropionic-N,N,N′,N′-tetraacetic acid (EDTACOOH), which is an EDTA derivative with a functionalisable carboxylic group, were used to coat NaGdF4. The carboxylic group was used to insert a polyethylene glycol (PEG) functionality (to give EDTAPEG) to favour better suspension and stealth and/or a fluorescent dye to obtain dual optical/MRI probes. The relaxometric behaviour of these NPs was investigated as a function of the magnetic field strength, and a significant contribution from water molecules hydrogen bonded to the organic coating was evidenced. The chemical stability of the NPs was evaluated both in a physiological medium and in the presence of a strong chelating agent. Finally, the dye-functionalised NPs were tested in ovarian carcinoma cells as dual optical and MRI probes.
Neuroblastoma is the most common type of cancer in infants. In children this tumor is particularl... more Neuroblastoma is the most common type of cancer in infants. In children this tumor is particularly aggressive; despite various new therapeutic approaches, it is associated with poor prognosis. Given the importance of endosomal-lysosomal proteolysis in cellular metabolism, we hypothesized that inhibition of lysosomal protease would impact negatively on neuroblastoma cell survival. Treatment with E-64 or CA074Me (2 specific inhibitors of cathepsin B) or with pepstatin A (a specific inhibitor of cathepsin D) was cytotoxic for 2 neuroblastoma cell lines having different degrees of malignancy. Cell death was associated with condensation and fragmentation of chromatin and externalization of plasma membrane phosphatidylserine, 2 hallmarks of apoptosis. Concomitant inhibition of the caspase cascade protected neuroblastoma cells from cathepsin inhibitor-induced cytotoxicity. These data indicate that prolonged inhibition of the lysosomal proteolytic pathway is incompatible with cell survival, leading to apoptosis of neuroblastoma cells, and that the cathepsin-mediated and caspase-mediated proteolytic systems are connected and cooperate in the regulation of such an event. Since modern antitumor chemotherapy is aimed at restoring the normal rate of apoptosis in neoplastic tissues, the demonstration that endosomal-lysosomal cathepsins are involved in this process may constitute a basis for novel strategies that include cathepsin inhibitors in the therapeutic regimen.
Hydrogen peroxide, the major oxidoradical species in the central nervous system, has been involve... more Hydrogen peroxide, the major oxidoradical species in the central nervous system, has been involved in neuronal cell death and associated neurodegenerative diseases. In this study, we have investigated the involvement of the lysosomal pathway in the cytotoxic mechanism of hydrogen peroxide in human neuroblastoma cells. Alteration of lysosomal and mitochondrial membrane integrity was shown to be an early event in the lethal cascade triggered by oxidative stress. Desferrioxamine (DFO), an iron chelator that abolishes the formation of reactive oxygen species within lysosomes, prevented lysosome leakage, mitochondrial permeabilization and caspase-dependent apoptosis in hydrogen peroxide-treated cells. Inhibition of cathepsin D, not of cathepsin B, as well as small-interference RNA-mediated silencing of the cathepsin D gene prevented hydrogen peroxide-induced injury of mitochondria, caspase activation, and TUNEL-positive cell death. Cathepsin D activity was shown indispensable for translocation of Bax onto mitochondrial membrane associated with oxidative stress. DFO abolished both the cytosolic relocation of Cathepsin D and the mitochondrial relocation of Bax in hydrogen peroxide-treated cells. siRNA-mediated down-regulation of Bax expression protected the cells from oxidoradical injury. The present study identifies the lysosome as the primary target and the axis cathepsin D-Bax as the effective pathway of hydrogen peroxide lethal activity in neuroblastoma cells.
The conference aims to discuss the latest advances in Cancer Sciences and establish collaborative... more The conference aims to discuss the latest advances in Cancer Sciences and establish collaborative networking. We invite senior scientists to join and share their knowledge with colleagues and young researchers. We encourage young researchers to take this opportunity to present their data and benefit from advice and constructive criticism in an open context.
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Papers by Ciro Isidoro